EBV noncoding RNA binds nascent RNA to drive host PAX5 to viral DNA

Authors:
Lee N, Moss WN, Yario TA, Steitz JA
In:
Source: Cell
Publication Date: (2015)
Issue: 160(4): 607-18
Research Area:
Immunotherapy / Hematology
Basic Research
Cells used in publication:
BJAB
Species: human
Tissue Origin: blood
Platform:
Nucleofector™ I/II/2b
4D-Nucleofector™ X-Unit
Experiment
2.5×106 BJAB-B1 cells were nucleofected with 10 µl of 100 mM KD-ASO/AMO stock solution in SF solution with program EN-150 using the Lonza 4D-Nucleofector System. HH514-16 cells were nucleofected using the Lonza 2b Device with solution V and program A-023. On the day following nucleofection, cells were separated from debris using Lymphocyte Separation Medium (Corning Cellgro) according to the manufacturer’s instructions.
Abstract
EBER2 is an abundant nuclear noncoding RNA expressed by the Epstein-Barr virus (EBV). Probing its possible chromatin localization by CHART revealed EBER2's presence at the terminal repeats (TRs) of the latent EBV genome, overlapping previously identified binding sites for the B cell transcription factor PAX5. EBER2 interacts with PAX5 and is required for the localization of PAX5 to the TRs. EBER2 knockdown phenocopies PAX5 depletion in upregulating the expression of LMP2A/B and LMP1, genes nearest the TRs. Knockdown of EBER2 also decreases EBV lytic replication, underscoring the essential role of the TRs in viral replication. Recruitment of the EBER2-PAX5 complex is mediated by base-pairing between EBER2 and nascent transcripts from the TR locus. The interaction is evolutionarily conserved in the related primate herpesvirus CeHV15 despite great sequence divergence. Using base-pairing with nascent RNA to guide an interacting transcription factor to its DNA target site is a previously undescribed function for a trans-acting noncoding RNA.